Redox sensitive cysteine residues in calbindin D(28)k are structurally and functionally important
(2005) In Biochemistry 44(2). p.684-693- Abstract
- Human calbindin D-28k is a Ca2+ binding protein that has been implicated in the protection of cells against apoptosis. In this study, the structural and functional significance of the five cysteine residues present in this protein have been investigated through a series of cystein e-to-serine mutations. The mutants were studied under relevant physiological redox potentials in which conformational changes were monitored using ANS binding. Urea-induced denaturations, as monitored by intrinsic tryptophan fluorescence, were also carried out to compare their relative stability. It was shown that the two N-terminal cysteine residues undergo a redox-driven structural change consistent with disulfide bond formation. The other cysteine residues are... (More)
- Human calbindin D-28k is a Ca2+ binding protein that has been implicated in the protection of cells against apoptosis. In this study, the structural and functional significance of the five cysteine residues present in this protein have been investigated through a series of cystein e-to-serine mutations. The mutants were studied under relevant physiological redox potentials in which conformational changes were monitored using ANS binding. Urea-induced denaturations, as monitored by intrinsic tryptophan fluorescence, were also carried out to compare their relative stability. It was shown that the two N-terminal cysteine residues undergo a redox-driven structural change consistent with disulfide bond formation. The other cysteine residues are not by themselves sufficient at inducing structural change, but they accentuate the disulfide-dependent conformational change in a redox-dependent manner. Mass spectrometry data show that the three C-terminal cysteine residues can be modified by glutathione. Furthermore, under oxidizing conditions, the data display additional species consistent with the conversion of cysteine thiols to sulfenic acids and disulfides to disulfide-S-monoxides. The biological function of calbindin D-28k appears to be tied to the redox state of the cysteine residues. The two N-terminal cysteine residues are required for activation of myo-inositol monophosphatase, and enzyme activation is enhanced under conditions in which these residues are oxidized. Last, oxidized calbindin D-28k binds Ca2+ with lower affinity than does the reduced protein. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/151574
- author
- Cedervall, Tommy LU ; Berggård, Tord LU ; Borek, V ; Thulin, Eva LU ; Linse, Sara LU and Akerfeldt, KS
- organization
- publishing date
- 2005
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Biochemistry
- volume
- 44
- issue
- 2
- pages
- 684 - 693
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- wos:000226348000027
- pmid:15641794
- scopus:12144274839
- ISSN
- 0006-2960
- DOI
- 10.1021/bi049232r
- language
- English
- LU publication?
- yes
- id
- 82de11d1-4aaa-4920-ac5c-a31df6a83423 (old id 151574)
- date added to LUP
- 2016-04-01 11:59:27
- date last changed
- 2022-01-26 21:11:47
@article{82de11d1-4aaa-4920-ac5c-a31df6a83423,
abstract = {{Human calbindin D-28k is a Ca2+ binding protein that has been implicated in the protection of cells against apoptosis. In this study, the structural and functional significance of the five cysteine residues present in this protein have been investigated through a series of cystein e-to-serine mutations. The mutants were studied under relevant physiological redox potentials in which conformational changes were monitored using ANS binding. Urea-induced denaturations, as monitored by intrinsic tryptophan fluorescence, were also carried out to compare their relative stability. It was shown that the two N-terminal cysteine residues undergo a redox-driven structural change consistent with disulfide bond formation. The other cysteine residues are not by themselves sufficient at inducing structural change, but they accentuate the disulfide-dependent conformational change in a redox-dependent manner. Mass spectrometry data show that the three C-terminal cysteine residues can be modified by glutathione. Furthermore, under oxidizing conditions, the data display additional species consistent with the conversion of cysteine thiols to sulfenic acids and disulfides to disulfide-S-monoxides. The biological function of calbindin D-28k appears to be tied to the redox state of the cysteine residues. The two N-terminal cysteine residues are required for activation of myo-inositol monophosphatase, and enzyme activation is enhanced under conditions in which these residues are oxidized. Last, oxidized calbindin D-28k binds Ca2+ with lower affinity than does the reduced protein.}},
author = {{Cedervall, Tommy and Berggård, Tord and Borek, V and Thulin, Eva and Linse, Sara and Akerfeldt, KS}},
issn = {{0006-2960}},
language = {{eng}},
number = {{2}},
pages = {{684--693}},
publisher = {{The American Chemical Society (ACS)}},
series = {{Biochemistry}},
title = {{Redox sensitive cysteine residues in calbindin D(28)k are structurally and functionally important}},
url = {{http://dx.doi.org/10.1021/bi049232r}},
doi = {{10.1021/bi049232r}},
volume = {{44}},
year = {{2005}},
}